Increased Cytotoxicity of Herpes Simplex Virus Thymidine Kinase Expression in Human Induced Pluripotent Stem Cells
| Autor: | Masahiro Toda, Hiroyuki Miyoshi, Chizuru Iwasawa, Ryota Tamura, Naoko Kuzumaki, Makoto Suematsu, Masaya Nakamura, Hideyuki Okano, Sadafumi Suzuki, Yuki Sugiura, Kazunari Yoshida, Minoru Narita |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Gene Expression Regulation
Viral induced pluripotent stem cells Genetic Vectors Apoptosis Biology nucleotide metabolism medicine.disease_cause Thymidine Kinase Catalysis Article Gene Expression Regulation Enzymologic Viral vector Inorganic Chemistry lcsh:Chemistry chemistry.chemical_compound cytotoxic medicine Gene silencing genome editing Humans Simplexvirus Clustered Regularly Interspaced Short Palindromic Repeats Physical and Theoretical Chemistry Induced pluripotent stem cell Molecular Biology lcsh:QH301-705.5 Ganciclovir Spectroscopy Thymidine triphosphate herpes simplex virus type 1 thymidine kinase Regulation of gene expression Gene Editing Nucleotides Organic Chemistry lentiviral vector Lentivirus Genes Transgenic Suicide General Medicine Genetic Therapy Suicide gene Computer Science Applications Cell biology Herpes simplex virus chemistry lcsh:Biology (General) lcsh:QD1-999 Thymidine kinase CRISPR-Cas Systems |
| Zdroj: | International Journal of Molecular Sciences International Journal of Molecular Sciences, Vol 20, Iss 4, p 810 (2019) Volume 20 Issue 4 |
| ISSN: | 1422-0067 |
| Popis: | Human induced pluripotent stem cells (iPSCs) hold enormous promise for regenerative medicine. The major safety concern is the tumorigenicity of transplanted cells derived from iPSCs. A potential solution would be to introduce a suicide gene into iPSCs as a safety switch. The herpes simplex virus type 1 thymidine kinase (HSV-TK) gene, in combination with ganciclovir, is the most widely used enzyme/prodrug suicide system from basic research to clinical applications. In the present study, we attempted to establish human iPSCs that stably expressed HSV-TK with either lentiviral vectors or CRISPR/Cas9-mediated genome editing. However, this task was difficult to achieve, because high-level and/or constitutive expression of HSV-TK resulted in the induction of cell death or silencing of HSV-TK expression. A nucleotide metabolism analysis suggested that excessive accumulation of thymidine triphosphate, caused by HSV-TK expression, resulted in an imbalance in the dNTP pools. This unbalanced state led to DNA synthesis inhibition and cell death in a process similar to a &ldquo thymidine block&rdquo but more severe. We also demonstrated that the Tet-inducible system was a feasible solution for overcoming the cytotoxicity of HSV-TK expression. Our results provided a warning against using the HSV-TK gene in human iPSCs, particularly in clinical applications. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|